Adhesion molecules expression analysis.

<p>(A) Relative mRNA levels of VCAM-1, ICAM-1, e-selectin and p-selectin in aortic arches from p55^+/+LDLR^−/− (n = 10) and p55^−/−LDLR^−/− (n = 8) mice. Values are represented relative to expression in p55^+/+LDLR^−/− arches. (B) Immunohistochemical staining of VCAM-1 expression on sections from aortic valve areas indicating a less intense endothelial staining in p55^−/−LDLR^−/− mice. Original magnification×200. (C) Staining quantification. * p = 0.01 by Student's t-test. Error bars indicate SEM.</p

Survival curves, aneurismal incidence and suprarenal aortic diameters.

<p>(A) Percent survival in p55^+/+LDLR^−/− (squares) and p55^−/−LDLR^−/− (triangles) mice following infusion with saline (n = 3 mice/group) or AngII (n = 13–14 mice/group). Comparison of survival curves between AngII-infused p55^+/+LDLR^−/− and p55^−/−LDLR^−/− mice gave a probability value of p = 0.054 by Pearson's chi-square test. (B) representative cross-sections of EVG stained suprarenal aortas from (a) saline infused and of advanced AAA in (b) p55^+/+LDLR^−/− and (c) p55^−/−LDLR^−/− mice indicating dissecting aneurysms and formation of thrombi. Original magnification×100 (a) and×50 (b,c). (C) Percent incidence of AAA in p55^+/+LDLR^−/− and p55^−/−LDLR^−/− mice (p = 0.2 by Fisher's exact test). (D) Suprarenal aortic diameters in p55^+/+LDLR^−/− (n = 12) and p55^−/−LDLR^−/− (n = 8) mice (p = 0.3 by Student's t-test).</p

General characterization of AngII infused mice.

<p>(A) Body weight (B) plasma cholesterol and (C) plasma triglyceride levels in p55^+/+LDLR^−/− and p55^−/−LDLR^−/− mice before (n = 13–14 mice/group) and after 5 weeks of high fat feeding (4 weeks of AngII infusion; n = 8–12 mice/group). (D) Plasma levels of pro-inflammatory cytokines and chemokines (n = 7–9 mice/group) after 5 weeks of high fat feeding (4 weeks of AngII infusion). * p<0.05 by Student's t-test. Error bars indicate SEM.</p

Body weight and plasma lipid levels.

<p>(A) Body weight (B) plasma cholesterol and (C) plasma triglyceride levels in p55^+/+LDLR^−/− (n = 18) and p55^−/−LDLR^−/− (n = 16) mice before and after 8 weeks of high fat feeding.</p

Cytokine and chemokine expression analysis.

<p>(A) Relative mRNA levels of IκBα, TNF, IL-6, IL-10 and (B) MCP-1, MIP-1α, MIP-1β, RANTES in aortic arches from p55^+/+LDLR^−/− (n = 10) and p55^−/−LDLR^−/− (n = 8) mice. Values are represented relative to expression in p55^+/+LDLR^−/− arches. (C) Plasma levels of pro-inflammatory cytokines and chemokines (n = 12–15 mice/group) after 8 weeks of high fat feeding. Error bars indicate SEM.</p

Atherosclerosis quantification.

<p>(A) Atherosclerotic lesion area in the aortic sinuses of p55^+/+LDLR^−/− (squares, n = 18) and p55^−/−LDLR^−/− (triangles, n = 16) mice. Each symbol represents one animal; bars represent means. * p = 0.02 by Student's t-test. (B) Representative lesions from p55^+/+LDLR^−/− and p55^−/−LDLR^−/− mice are shown. Original magnification×40. (C) Lesion classification according to severity. (D) Gene expression analysis in p55^+/+LDLR^−/− (n = 10) and p55^−/−LDLR^−/− (n = 8) aortic arches. Values are represented relative to expression in p55^+/+LDLR^−/− arches. * p = 0.03 by Student's t-test. Error bars indicate SEM.</p

Presentation_1_Intracrine Regulation of Estrogen and Other Sex Steroid Levels in Endometrium and Non-gynecological Tissues; Pathology, Physiology, and Drug Discovery.pdf

Our understanding of the intracrine (or local) regulation of estrogen and other steroid synthesis and degradation expanded in the last decades, also thanks to recent technological advances in chromatography mass-spectrometry. Estrogen responsive tissues and organs are not passive receivers of the pool of steroids present in the blood but they can actively modify the intra-tissue steroid concentrations. This allows fine-tuning the exposure of responsive tissues and organs to estrogens and other steroids in order to best respond to the physiological needs of each specific organ. Deviations in such intracrine control can lead to unbalanced steroid hormone exposure and disturbances. Through a systematic bibliographic search on the expression of the intracrine enzymes in various tissues, this review gives an up-to-date view of the intracrine estrogen metabolisms, and to a lesser extent that of progestogens and androgens, in the lower female genital tract, including the physiological control of endometrial functions, receptivity, menopausal status and related pathological conditions. An overview of the intracrine regulation in extra gynecological tissues such as the lungs, gastrointestinal tract, brain, colon and bone is given. Current therapeutic approaches aimed at interfering with these metabolisms and future perspectives are discussed.</p

General reduction of diet-induced hepatic inflammation in <i>LDLR</i>^−/−/<i>MPO</i>^−/−tp mice.

<p>A) Significantly lower number of hepatic Ly-6G⁺ neutrophils and CD3⁺ T-lymphocytes in <i>LDLR</i>^−/−/<i>MPO</i>^−/−tp mice as compared with <i>LDLR</i>^−/−/<i>MPO</i>^+/+tp mice after 8 weeks of high-fat feeding (Ly-6G: 36.7±2.6 vs. 47.8±3.1 cells/mm², p = 0.03; CD3: 49.1±4.2 vs. 62.9±5.0 cells/mm², p = 0.04). Pictures represent examples of the stainings (200× magnification). B) Hepatic pro-inflammatory cytokine/chemokine expression is substantially reduced in <i>LDLR</i>^−/−/<i>MPO</i>^−/−tp mice after 8 weeks high-fat diet (TNF-α 1.8-fold, p = 0.03, IL-1α 1.6-fold, p<0.01, IL-6 1.3-fold, p = 0.67, Mcp-1 2.5-fold, p<0.01), in parallel with a reduction of CD68 expression (1.3-fold, p<0.05).</p

Wound Administration of M2-Polarized Macrophages Does Not Improve Murine Cutaneous Healing Responses

<div><p>Macrophages play a crucial role in all stages of cutaneous wound healing responses and dysregulation of macrophage function can result in derailed wound repair. The phenotype of macrophages is influenced by the wound microenvironment and evolves during healing from a more pro-inflammatory (M1) profile in early stages, to a less inflammatory pro-healing (M2) phenotype in later stages of repair. The aim of the current study was to investigate the potential of exogenous administration of M2 macrophages to promote wound healing in an experimental mouse model of cutaneous injury. Bone marrow derived macrophages were stimulated in-vitro with IL-4 or IL-10 to obtain two different subsets of M2-polarized cells, M2a or M2c respectively. Polarized macrophages were injected into full-thickness excisional skin wounds of either C57BL/6 or diabetic db/db mice. Control groups were injected with non-polarized (M0) macrophages or saline. Our data indicate that despite M2 macrophages exhibit an anti-inflammatory phenotype in-vitro, they do not improve wound closure in wild type mice while they delay healing in diabetic mice. Examination of wounds on day 15 post-injury indicated delayed re-epithelialization and persistence of neutrophils in M2 macrophage treated diabetic wounds. Therefore, topical application of ex-vivo generated M2 macrophages is not beneficial and contraindicated for cell therapy of skin wounds.</p></div

Decreased cholesterol accumulation in the liver of <i>LDLR</i>^−/−/<i>MPO</i>^−/−tp mice.

<p>A) Representative Oil red O stainings of liver sections of <i>LDLR</i>^−/−/<i>MPO</i>^−/−tp and <i>LDLR</i>^−/−/<i>MPO</i>^+/+tp mice fed a high-fat diet for 8 weeks, showing comparable lipid accumulation (100× magnification). B) Similar hepatic triglyceride levels in <i>LDLR</i>^−/−/<i>MPO</i>^−/−tp and <i>LDLR</i>^−/−/<i>MPO</i>^+/+tp mice after high-fat feeding (0.31±0.02 vs. 0.35±0.02 µg/µg protein, p = 0.24). Chow-fed <i>LDLR</i>^−/−/<i>MPO</i>^+/+ mice show a lower level of liver triglycerides. C) Plasma triglyceride levels are similar in <i>LDLR</i>^−/−/<i>MPO</i>^−/−tp and <i>LDLR</i>^−/−/<i>MPO</i>^+/+tp animals after high-fat feeding (1.50±0.09 vs. 1.64±0.09 mmol/l, p = 0.42). D) High-fat feeding results in higher plasma cholesterol levels in <i>LDLR</i>^−/−/<i>MPO</i>^+/+tp animals as compared with <i>LDLR</i>^−/−/<i>MPO</i>^−/−tp mice (33.5±0.1 vs. 39.5±2.0 mmol/l, p = 0.02). E) Diet-induced liver cholesterol accumulation is reduced in <i>LDLR</i>^−/−/<i>MPO</i>^−/−tp mice compared with <i>LDLR</i>^−/−/<i>MPO</i>^+/+tp animals (0.072±0.004 vs. 0.090±0.004 µg/µg protein, p = 0.01), but does not reach the level observed in chow-fed <i>LDLR</i>^−/−/<i>MPO</i>^+/+ mice. F) Hepatic mRNA expression of key enzymes in cholesterol metabolism is not altered in <i>LDLR</i>^−/−/<i>MPO</i>^−/−tp mice, whereas scavenger receptor expression is reduced (SR-B1 1.7-fold, p<0.01; CD36 1.4-fold, p<0.01, SR-A 1.2-fold, p = 0.63).</p